ABSTRACT
Side versus forward light scattergrams, and fluorescence (488 nm excitation) intensity versus particle count histograms were gathered for bare, R6G-coated, and silica-R6G-coated silver particles of 150-200 nm diameter, one-by-one by flow cytometry. Fluorescence emission intensity of the composite particles monotonically increased and then reached a plateau with greater R6G concentrations, as measured by flow cytometry. Fluorescence amplification factors of up to 3.5x10(3) were estimated by reference to measurements on core-shell particles with silica instead of silver cores. Huge surface enhanced Raman scattering (SERS) intensities, at least 10(14)-fold greater than normal Raman scattering intensities, were observed with 633 nm excitation for molecules such as rhodamine 6G (R6G) on the same single particles of silver. Although routine transmission (TEM) and scanning (SEM) electron microscopies showed gross structures of the bare and coated particles, high-resolution field emission scanning electron microscopy (FE-SEM), revealed Brownian roughness describing quantum size and larger structures on the surface of primary colloidal silver particles. These silver particles were further characterized by extinction spectra and zeta potentials. Structural and light scattering observations that are reported herein were used to tentatively propose a new hierarchical model for the mechanism of SERS.
ABSTRACT
Water-dispersible Amdex-CdSe nanoparticle complexes with sufficient luminescence intensity were prepared at room temperature by rapidly mixing aqueous solutions of either sodium selenide or selenourea with those of cadmium chloride in the presence of amino-derivatized polysaccharides (Amdex) as stabilizing agent. It was shown that the size of CdSe crystallites decreased with increasing content of the polymer in the precipitation process. When present in a sufficient amount, Amdex was found to be an effective stabilizing and capping agent, producing CdSe nanocrystals of weak-to-medium luminescence intensity (maximum room temperature quantum yields of 15-16%). Furthermore, Amdex has proven to be an effective protective agent against photochemical degradation.
ABSTRACT
Uniform fluorescent silica-dye microspheres have been prepared by coating preformed monodispersed silica particles with silica layers containing rhodamine 6G or acridine orange. The resulting dispersions exhibit intense fluorescent emission between 500 and 600 nm, over a broad excitation wavelength range of 460 to 550 nm, even with exceedingly small amounts of dyes incorporated into the silica particles (10-30 ppm, expressed as weight of dye relative to weight of dry particles). The fluorescent particles can be prepared in micrometer diameters suitable for analyses using flow cytometry with 488-nm laser excitation.
ABSTRACT
CD4 or CD8 antibodies were covalently bound to latex beads by reaction of activated CD4 or CD8 monoclonal antibodies with 2-µm-diameter, 1,3-diaminopropane (DAP) coupled, polystyrene aldehyde/sulfate latex beads. Spectrophotometric analyses of the filtrates of the antibody-bead conjugation mixtures for unreacted antibody allowed construction of binding curves of antibody for the polystyrene bead surface and evaluation of binding constants for association of antibody with bead, ranging from 1.5x10(7) to 1.6x10(7) M(-1) for CD4 and CD8 antibodies. The reaction of the antibody thiol group with the activated maleimide group on the bead at pH 7.2-7.3 was complete within 10-15 min. The kinetics of CD4 or CD8 monoclonal antibody displacement from the surface of covalently conjugated antibody-polystyrene latex beads was followed as a function of temperature (5, 22, and 37 degrees C) and the nature of the final diluent for the antibody-coated beads by measuring the concentration of antibody in the filtrates of conjugated beads by an ELISA (enzyme-linked immunosorbent assay). The displacement reaction showed a pseudo-zero-order dependence of the rate, with constants, k(1), ranging from 0.65x10(-17) to 270x10(-17) M s(-1). The functionality of antibody-coated beads suspended in various media was also monitored in a biological cell assay with whole blood. The cell assay depends on forming a layer of beads around targeted lymphocytes to distinguish them from nontargeted lymphocytes by differences in dc or rf conductivity or median angle light scatter. Covalently bound CD4 and CD8 antibody beads stored in one set of media at 5, 22, and 37 degrees C over a period of 16 weeks showed excellent results in the STKS assay with various blood donors, which correlated well (correlation coefficients of 0.99 for CD4 data and 0.93 for CD8 data) with reference results obtained with fluorescent markers by flow cytometry. Covalently bound CD4/CD8 beads stored for 2 weeks in BSA buffer at 5-37 degrees C performed equally well in providing accurate values of the percentage of CD4- or CD8-positive cells in the total white blood cell population, whereas the same beads stored in the 47-50 degrees C range showed some failures in performance. Comparison with antibody concentrations in filtrates of adsorbed antibody-bead suspensions showed 2- to 10-fold greater amounts of free antibody at comparable elapsed time, media, and temperature conditions. A threshold of 1-2 µg/mL of free antibody was necessary before adverse effects on the biological cell assay were noticeable. Copyright 2001 Academic Press.
